Human trophoblast stem cells and use thereof

• US 8,497,120 B2
• Filed: 03/08/2012
• Issued: 07/30/2013
• Est. Priority Date: 02/24/2005
• Status: Active Grant

• Current Assignee: Accelerated Biosciences Corp.
• Original Assignee: Jau-Nan Lee, Tony Tung-Ying Lee, Yuta Lee
• Inventors: Lee, Jau-Nan, Lee, Tony Tung-Ying, Lee, Yuta
• Primary Examiner(s): Kelly, Robert M
• Application Number:
  US13/415,595
• Publication Number:
  US 20120328579A1
• Time in Patent Office: 509 Days
• Field of Search: None
• US Class Current: 435/325
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 35/54 Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0677 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: G01N 33/5073 Stem cells

Abstract

Existence of human trophoblast stem (hTS) cells has been suspected but unproved. The isolation of hTS cells is reported in the early stage of chorionic villi by expressions of FGF4, FGFR-2, Oct4, Thy-1, and stage-specific embryonic antigens distributed in different compartments of the cell. hTS cells are able to derive into specific cell phenotypes of the three primitive embryonic layers, produce chimeric reactions in mice, and retain a normal karyotype and telomere length. In hTS cells, Oct4 and fgfr-2 expressions can be knockdown by bFGF. These facts suggest that differentiation of the hTS cells play an important role in implantation and placentation. hTS cells could be apply to human cell differentiation and for gene and cell-based therapies.

Claims

1. A method for isolating a human trophoblast stem cell, comprising the steps of:

a. obtaining trophoblastic villi from a tubal ectopic pregnant mass;

b. collecting cells from the trophoblastic villi; and

c. culturing the collected cells in a culture medium to obtain the isolated human trophoblast stem cell, wherein the human trophoblast stem cell expresses Octamer-4(Oct4) and SSEA-4.

2. The method of claim 1 further comprising cutting the trophoblastic villi into pieces.

3. The method of claim 1 further comprising treating the trophoblastic villi with an enzyme.

4. The method of claim 1, wherein the human trophoblast stem cell is genetically modified to introduce a mutation into the cell.

5. The method of claim 1, wherein the pregnant mass is obtained in an unruptured manner.

6. The method of claim 1, wherein the pregnant mass is at a gestational age of no older than 7 or 8 weeks.

7. The method of claim 1, wherein the culture medium is free of a feeder layer.

8. The method of claim 1 further comprising the steps of:

a. forming embryonic bodies (EBs) in the culture medium;

b. treating the EBs with an enzyme; and

c. collecting cells from the enzyme-treated EBs to obtain the isolated human trophoblast stem cell.